Are bioplastics safe? Hazardous effects of polylactic acid (PLA) nanoplastics in Drosophila.

The expanded uses of bioplastics require understanding the potential health risks associated with their exposure. To address this issue, Drosophila melanogaster as a versatile terrestrial in vivo model was employed, and polylactic acid nanoplastics (PLA-NPLs), as a proxy for bioplastics, were tested as a material model. Effects were determined in larvae exposed for 4 days to different concentrations (25, 100, and 400 µg/mL) of 463.9 ± 129.4 nm PLA-NPLs. Transmission electron microscopy (TEM) and scanning electron microscope (SEM) approaches permitted the detection of PLA-NPLs in the midgut lumen of Drosophila larvae, interacting with symbiotic bacteria. Enzymatic vacuoles were observed as carriers, collecting PLA-NPLs and enabling the crossing of the peritrophic membrane, finally internalizing into enterocytes. Although no toxic effects were observed in egg-to-adult survival, cell uptake of PLA-NPLs causes cytological disturbances and the formation of large vacuoles. The translocation across the intestinal barrier was demonstrated by their presence in the hemolymph. PLA-NPL exposure triggered intestinal damage, oxidative stress, DNA damage, and inflammation responses, as evaluated via a wide set of marker genes. Collectively, these structural and molecular interferences caused by PLA-NPLs generated high levels of oxidative stress and DNA damage in the hemocytes of Drosophila larvae. The observed effects point out the need for further studies aiming to deepen the health risks of bioplastics before adopting their uses as a safe plastic alternative.

Titanium-doped PET nanoplastics, from opaque milk bottle degradation, as a model of environmental true-to-life nanoplastics. Hazardous effects on Drosophila.

Micro and nanoplastics (MNPLs) are emergent environmental pollutants, resulting from the degradation of plastic waste, requiring urgent information on their potential risks to human health. To determine such risks, reliable true-to-life materials are essential. In this work, we have used titanium-doped PET NPLs [PET(Ti)NPLs], obtained by grinding opaque milk polyethylene terephthalate (PET) bottles, as a true-to-life MNPLs model. These opaque PET bottles, with an average size of 112 nm, contain about 3% Ti in the form of titanium dioxide rod nanoparticles. TEM investigation confirmed the mixed Ti/PET nature of the obtained true-to-life NPLs, and the rod shape of the embedded TiO2NPs. In the in vivo Drosophila model neither PET(Ti)NPLs nor TiO2NPs reduced the survival rates, although their internalization was confirmed in different compartments of the larval body by using confocal and transmission electron microscopies. The presence of Ti in the PET(Ti)NPLs permitted to quantify its presence both in larvae (2.1 ± 2.2 µg/g of Ti) and in the resulting adults (3.4 ± 3.2 µg/g of Ti) after treatment with 500 µg/g food of PET(Ti)NPL, suggesting its potential use to track their fate in more complex organisms such as mammals. PET(Ti)NPLs, as well as TiO2NPs, altered the expression of genes driving different response pathways, inducing significant oxidative stress levels (up to 10 folds), and genotoxicity. This last result on the genotoxic effects is remarkable in the frame of the hot topic discussion on the risk that titanium compounds, used as food additives, may pose to humans.

Melatonin downregulates the increased hepatic alpha-fetoprotein expression and restores pancreatic beta cells in a streptozotocin-induced diabetic rat model: a clinical, biochemical, immunohistochemical, and descriptive histopathological study.

Background: Diabetes mellitus (DM) is a chronic metabolic disorder. Hepatopathy is one of the serious effects of DM Melatonin (MT) is a potent endogenous antioxidant that can control insulin output. However, little information is available about the potential association between melatonin and hepatic alpha-fetoprotein expression in diabetes. Objective: This study was conducted to assess the influence of MT on diabetes-related hepatic injuries and to determine how ß-cells of the pancreas in diabetic rats respond to MT administration. Materials and methods: Forty rats were assigned to four groups at random (ten animals per group). Group I served as a normal control group. Group II was induced with DM, and a single dose of freshly prepared streptozotocin (45 mg/kg body weight) was intraperitoneally injected. In Group III, rats received 10 mg/kg/day of intraperitoneal melatonin (IP MT) intraperitoneally over a period of 4 weeks. In Group IV (DM + MT), following the induction of diabetes, rats received MT (the same as in Group III). Fasting blood sugar, glycosylated hemoglobin (HbA1c), and serum insulin levels were assessed at the end of the experimental period. Serum liver function tests were performed. The pancreas and liver were examined histopathologically and immunohistochemically for insulin and alpha-fetoprotein (AFP) antibodies, respectively. Results: MT was found to significantly modulate the raised blood glucose, HbA1c, and insulin levels induced by diabetes, as well as the decreased alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Furthermore, MT attenuated diabetic degenerative changes in the pancreas and the hepatic histological structure, increased the ß-cell percentage area, and decreased AFP expression in the liver tissue. It attenuated diabetes-induced hepatic injury by restoring pancreatic ß-cells; its antioxidant effect also reduced hepatocyte injury. Conclusion: Collectively, the present study confirmed the potential benefits of MT in downregulating the increased hepatic alpha-fetoprotein expression and in restoring pancreatic ß-cells in a streptozotocin-induced diabetic rat model, suggesting its promising role in the treatment of diabetes.

The hazardous impact of true-to-life PET nanoplastics in Drosophila.

Plastic pollution is a continuously growing problem that can threaten wildlife and human beings. Environmental plastic waste is degraded into small particles termed micro/ nanoplastics (MNPLs) that, due to their small size, can be easily internalized into the exposed organisms, increasing the risks associated with their exposure. To appropriately determine the associated health risk, it is essential to obtain/test representative MNPLs' environmental samples. To such end, we have obtained NPLs resulting from sanding commercial water polyethylene terephthalate (PET) bottles. These true-to-life PETNPLs were extensively characterized, and their potential hazard impacts were explored using Drosophila melanogaster. To highlight the internalization through the digestive tract and the whole body, transmission electron microscopy (TEM) and confocal microscopy were used. In spite of the observed efficient uptake of PETNPLs into symbiotic bacteria, enterocytes, and hemocytes, the exposure failed to reduce flies' survival rates. Nevertheless, PETNPLs exposure disturbed the expression of stress, antioxidant, and DNA repair genes, as well as in those genes involved in the response to physical intestinal damage. Importantly, both oxidative stress and DNA damage induction were markedly increased as a consequence of the exposure to PETNPLs.

Antagonistic in vivo interaction of polystyrene nanoplastics and silver compounds. A study using Drosophila.

Since heavy metals and micro-/nanoplastics (MNPLs) can share common environmental niches, their potential interactions could modulate their hazard impacts. The current study was planned to evaluate the potential interactions between silver compounds (silver nanoparticles or silver nitrate) and two different sizes of polystyrene nanoplastics (PSNPLs) (PS-50 and PS-500 nm), administered via ingestion to Drosophila larvae. While egg-to-adult survival was not affected by the exposure to silver compounds, PSNPLs, or their coexposures, the combined treatments succeeded to restore the delay of fly emergence induced by silver compounds. Transmission electron microscopy (TEM) and inductively coupled plasma mass spectrometry (ICP-MS) showed the ability of PSNPLs to transport silver compounds (regardless of their form) across the intestinal barrier, delivering them into the hemolymph of Drosophila larvae in a concentration exceeding that mediated by the exposure to silver compounds alone. The molecular response (gene expression) of Drosophila larvae greatly fluctuated, accordingly if exposures were administered alone or in combination. Although PSNPLs produced some oxidative stress in the hemocytes of Drosophila, especially at the highest dose (1 mM), higher levels were observed after silver exposure, regardless of its form. Interestingly, the oxidative stress of silver, especially that produced by nano­silver, drastically decreased when coexposed with PSNPLs. Similar effects were observed regarding the DNA damage induced in Drosophila hemocytes, where cotreatment decreased the genotoxicity induced by silver compounds. This antagonistic interaction could be attributed to the ability of tiny plastic specks to confine silver, avoiding its bioavailability, and diminishing their potential impacts.

Spontaneous changes in brain striatal dopamine synthesis and storage dynamics ex vivo reveal end-product feedback-inhibition of tyrosine hydroxylase.

Synaptic events are important to define treatment strategies for brain disorders. In the present paper, freshly obtained rat brain striatal minces were incubated under different times and conditions to determine dopamine biosynthesis, storage, and tyrosine hydroxylase phosphorylation. Remarkably, we found that endogenous dopamine spontaneously accumulated during tissue incubation at 37 °C ex vivo while dopamine synthesis simultaneously decreased. We analyzed whether these changes in brain dopamine biosynthesis and storage were linked to dopamine feedback inhibition of its synthesis-limiting enzyme tyrosine hydroxylase. The aromatic-l-amino-acid decarboxylase inhibitor NSD-1015 prevented both effects. As expected, dopamine accumulation was increased with l-DOPA addition or VMAT2-overexpression, and dopamine synthesis decreased further with added dopamine, the VMAT2 inhibitor tetrabenazine or D2 auto-receptor activation with quinpirole, accordingly to the known synaptic effects of these treatments. Phosphorylation activation and inhibition of tyrosine hydroxylase on Ser31 and Ser40 with okadaic acid, Sp-cAMP and PD98059 also exerted the expected effects. However, no clear-cut association was found between dopamine feedback inhibition of its own biosynthesis and changes of tyrosine hydroxylase phosphorylation, assessed by Western blot and mass spectrometry. The later technique also revealed a new Thr30 phosphorylation in rat tyrosine hydroxylase. Our methodological assessment of brain dopamine synthesis and storage dynamics ex vivo could be applied to predict the in vivo effects of pharmacological interventions in animal models of dopamine-related disorders.